Pitfalls of quantitative real-time reverse-transcription polymerase chain reaction.

By Stephen A. Bustin, T. Nolan. Journal of Biomolecular Techniques, 2004

1,138 citations recorded by the source corpus.

View Paper: https://www.semanticscholar.org/paper/62dbed87fd0aa453f3db5068711288fd2cc2168d

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Citations in the corpus

Quantitative real-time RT-PCR--a perspective. (2005, methodology): “The C t is used for quantifying target copy number, yet its value is entirely subjective, as the threshold can be altered at will (Bustin & Nolan 2004).”
Identification of suitable reference genes for studying gene expression in cucumber plants subjected to abiotic stress and growth regulators (2011, background): “The main advantages of this technique are high sensitivity, high specificity and broad quantification range (Bustin et al. 2005; Bustin and Nolan 2004; Gahon et al. 2004; Ginzinger 2002).”
Quantitative real-time RT-PCR data analysis: current concepts and the novel “gene expression’s CT difference” formula (2006, background): “Several factors, such as phenol, ethanol, haemoglobin, heparin and even the reverse transcriptase, are known to inhibit PCR efficiency [22, 23].”
Quantitative and stoichiometric analysis of the microRNA content of exosomes (2014, background): “Third, we addressed the possibility that our results could be the product of inaccurate miRNA quantification caused by the dependence of real-time PCR on standard curves (37).”
Droplet Digital PCR versus qPCR for gene expression analysis with low abundant targets: from variable nonsense to publication quality data (2017, background): “The most susceptible samples include those with high levels of impurities and low target concentrations because these samples cannot be adequately diluted to eliminate the effect of contamination on data quality 1, 21 .”
The implications of using an inappropriate reference gene for real-time reverse transcription PCR data normalization. (2005, background): “It is now clear, however, that in most experimental situations the use of conventional reference genes, such as glyceraldehyde-3-phosphate dehydrogenase (GAPDH) and -actin, is inappropriate due to their variability [1,2].”
The MIQE guidelines: minimum information for publication of quantitative real-time PCR experiments. (2009, background): “This remarkable lack of consensus on how best to perform qPCR experiments has the adverse consequence of perpetuatingastringofseriousshortcomingsthatencumber its status as an independent yardstick ( 12 ).”
Comprehending crystalline β-carotene accumulation by comparing engineered cell models and the natural carotenoid-rich system of citrus (2011, unknown): “It is inappropriate to compare degraded and intact samples and thus necessitates a systematic RNA integrity control prior to any qPCR analysis (Bustin and Nolan, 2004; Imbeaud et al., 2005; PérezNovo et al., 2005).”
Standard methods for virus research in Apis mellifera (2013, background): “They are also much cheaper to design, adapt and optimize (Bustin and Nolan, 2004).”
Standard methods for molecular research in Apis mellifera (2013, background): “As dsRNAs are processed by the dicer complex into a cocktail of siRNAs 19–21 nt in length, the absence of 20-nt stretches of homology minimizes the possibility of off-target effects.”
Comparison of relative mRNA quantification models and the impact of RNA integrity in quantitative real-time RT-PCR (2006, background): “With regard to the efficiency of the PCR, those can affect by a number of variables like length of the amplicon, RNA secondary structure and primer quality (Bustin and Nolan 2004; Wong and Medrano 2005 ).”
RNA integrity and the effect on the real-time qRT-PCR performance. (2006, methodology): “5, some, low-throughput method and requires significant amounts of precious RNA (Bustin and Nolan, 2004a).”